Precise tumor immune rewiring via synthetic CRISPRa circuits gated by concurrent gain/loss of transcription factors

Abstract

Reinvigoration of antitumor immunity has recently become the central theme for the development of cancer therapies. Nevertheless, the precise delivery of immunotherapeutic activities to the tumors remains challenging. Here, we explore a synthetic gene circuit-based strategy for specific tumor identification, and for subsequently engaging immune activation. By design, these circuits are assembled from two interactive modules, i.e., an oncogenic TF-driven CRISPRa effector, and a corresponding p53-inducible off-switch (NOT gate), which jointly execute an AND-NOT logic for accurate tumor targeting. In particular, two forms of the NOT gate are developed, via the use of an inhibitory sgRNA or an anti-CRISPR protein, with the second form showing a superior performance in gating CRISPRa by p53 loss. Functionally, the optimized AND-NOT logic circuit can empower a highly specific and effective tumor recognition/immune rewiring axis, leading to therapeutic effects in vivo. Taken together, our work presents an adaptable strategy for the development of precisely delivered immunotherapy.

CRISPR/Cas9-Mediated Genome Editing in Cancer Therapy

“…This methodology aids in identifying new resistance targets, enhancing clinical therapeutic opportunities. Wang et al [70] developed a strategy based on CRISPRa and synthetic gene circuits for the identification of specific tumors, followed by the activation of immune-related genes. This circuit consists of two interactive modules, namely the oncogenic TF-driven CRISPRa effector (AND gate) and the corresponding p53-induced shut-off switch (NOT gate), working together to implement accurate tumor targeting through AND-NOT logic.…”

In Situ Tumor Vaccine for Lymph Nodes Delivery and Cancer Therapy Based on Small Size Nanoadjuvant

“…Cancer immunotherapy can enhance the antitumor immune response to fight against tumor cells by activating the immune system, achieving a durable antitumor immune response and preventing tumor metastasis and recurrence, which has attracted the attention of many scientists and shown great promise in cancer treatment during the past decades. [ 1–3 ] Various kinds of cancer immunotherapeutic approaches, such as immune checkpoint blockade (ICB) therapy, [ 4–6 ] chimeric antigen receptor T‐cell (CAR‐T) therapy, [ 7,8 ] and tumor vaccines [ 9,10 ] have been extensively investigated with desirable outcomes. Among them, tumor vaccine, which can generate specific anti‐tumor immune response, is supposed to be one of the most important immunotherapeutic strategies and holds tremendous potential for cancer treatment.…”

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promise in cancer treatment during the past decades. [1][2][3] Various kinds of cancer immunotherapeutic approaches, such as immune checkpoint blockade (ICB) therapy, [4][5][6] chimeric antigen receptor T-cell (CAR-T) therapy, [7,8] and tumor vaccines [9,10] have been extensively investigated with desirable outcomes. Among them, tumor vaccine, which can generate specific anti-tumor immune response, is supposed to be one of the most important immunotherapeutic strategies and holds tremendous potential for cancer treatment.

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